Molecular Mechanism Of MT-12 Inhibiting Invasion And Metastasis Of Bladder Cancer Cells

Objectives:After MT-12 was applied to bladder cancer cells,the expression of cell invasion and metastasis-related factors was detected,and the molecular mechanism of MT-12 inhibiting invasion and metastasis of bladder cancer cells was explored.Methods:1.Apply different MT-12 concentrations to treat bladder cancer cells T24 and RT4;then use the tumor suppressor phorbol ester PMA to intervene in bladder cancer cells:divided into control group,PMA and PMA+MT-12 three groups;2.Gelatin zymography was used to detect the activity of MMP-2 and MMP-9 in T24 and RT4 cell supernatants under different MT-12 concentrations and PMA intervention;3.ELIS A was used to detect the expression of VEGF in T24 and RT4 cell supernatants under different MT-12 concentrations and PMA intervention;4.qRT-PCR was used to detect the expression of metastasis-associated factors ICAM-1,VCAM-1 and VEGF under different MT-12 and PMA interventions;5.Western Blot was used to detect the protein expression levels of NF-?B and EGFR/PI3K/Akt signaling pathways under MT-12 treatment.Results:l.The expression changes of MMP family were detected.The results showed that the activity of MMP-9 was significantly down-regulated and the activity of MMP-2 was not significantly changed after MT-12 treatment.MT-12 could effectively inhibit the up-regulation of MMP-9 induced by PMA.However,there was no significant effect on the activity of MMP-2.2.VEGF was detected in bladder cancer cells.ELISA and qRT-PCR results showed that PMA promoted the secretion of bladder cancer cells,while MT-12 inhibited the expression of VEGF.3.The related adhesion factors ICAM-1 and VCAM-1 were detected.The results of qRT-PCR showed that the expression of ICAM-1 was significantly decreased after treatment with different concentrations of MT-12,while the expression of VCAM-1 was not affected;PMA was up-regulated significantly.The level of ICAM-1,while the level of VCAM-1 was unchanged,MT-12 was able to inhibit the upregulation of ICAM-1,and had no effect on the expression of VCAM-1.4.Western Blot results showed that MT-12 can inhibit the activation of phosphorylated NF-?B p65,but not by affecting the EGFR/PI3K/Akt signaling pathway.Conclusions:MT-12 can inhibit the activity or expression of bladder cancer invasion and metastasis related factors MMP-9,VEGF and ICAM-1 by NF-?B signaling pathway in vitro,so as to inhibit the ability of bladder cancer cells to open the gap of endothelial cells.